Site-Directed Mutagenesis of Large Plasmids | Zendy

Susan A. NadinDavis | Zendy; Shirley C. Chang | Zendy

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Site-Directed Mutagenesis of Large Plasmids

Author(s) -

Susan A. NadinDavis,

Shirley C. Chang

Publication year - 1998

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/98256st04

Subject(s) - plasmid , mutagenesis , biology , mutant , site directed mutagenesis , genetics , mutation , insertional mutagenesis , overlap extension polymerase chain reaction , computational biology , vector (molecular biology) , recombinant dna , dna , gene

A protocol combining recombination PCR and long-distance PCR is demonstrated to be highly accurate and rapid for site-directed mutagenesis of large (> 10 kb) plasmids. Application of this protocol to the generation of mutant rabies virus glycoproteins expressed by the baculovirus/insect cell system illustrates the usefulness of this approach in facilitating structure-function relationships in this important eukaryotic expression system.

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